Hi!
I am trying to measure hemolysis in plasma and serums using one spectrophotometer(EPOCH) and Nanodrop at 414nm. I always obtain higher values using nanodrop comparing with the same sample at Spectrophotometer. does anyone know about it? and what is the best option to mesure the haemolysis in plasma and serums? I work with miRnas and it is very important for me! Thank you very much!
María
I hope that this publication will be helpful
https://www.researchgate.net/publication/299983454_Comparison_of_Methodologies_to_Detect_Low_Levels_of_Hemolysis_in_Serum_for_Accurate_Assessment_of_Serum_microRNAs
Article Comparison of Methodologies to Detect Low Levels of Hemolysi...
Some common paper, those might will help you.
http://www.biochemia-medica.com/content/hemolysis-detection-and-management-hemolyzed-specimens
http://clinchem.aaccjnls.org/content/53/9/1717
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4330168/
Dear Gregory and Dear Babu Mia:
Thank you very much for answering me! I am sure that these papers will be very useful for my work!
Thank you!
María
Dear Maria,
You can read this paper,
Hemolysis in diagnostic samples, Revista del Laboratorio Clínico, Rev Lab Clin. 2009;2(4):185–195.
In most cases, hemolysis is an avoidable preanalytical effect. It can appear as a result of the procedure used during blood specimen collection and also due to transport conditions and sample preparation. Hemolysis can lead to errors in many common determinations in clinical chemistry, mostly due to the leakage of cellular contents into the plasma, and in some cases, by the interfering effect of red blood cells components, such as hemoglobin. Laboratories need to be able to detect and measure hemolysis by a standardized procedure. The majority of current biochemistry analyzers can measure hemolysis by a spectrophotometric method. Nevertheless, the influence of hemolysis depends on the measurement method employed. Laboratories should demand that manufacturers give
detailed information and make exhaustive studies regarding the influence of hemolysis on each analyte. This would allow each laboratory to establish the degree of hemolysis that produces a significant error in an analysis and in accordance with the laboratory’s quality specifications.
Hi!Thank you very much for your help Daniel!!
Does anyone know anything about the measurement of bilirubin in plasma samples? Because I think that sometimes I obtain higher values of Absorbance at 414 nm,when I try to measure the hemolysis in plasma, due to the interference of Bilirrubine. If anyone knows something about this, I would be very grateful!
Thank you very much!!!
Hola Maria, Some spectroscopic methods conducted in 1980s may be helpful. For example, this paper that mentions Allen correction (PMID: 6713623). In my study, we made sure that the 'miR ratio' was equally distributed in all treatment groups. however, if you try to have equal distribution of Nanodrop readings in your treatment groups as a proxy for 'miR ratio', it MAY BE OK (a strong emphasis on MAY) since you are distributing miRs affected by haemolysis equally in all groups.
If you want to know what miRs are affected by haemolysis, search for papers that did Sequencing or PCR on miRs present in RBC. For example, See Table 5 in Kirshner's paper: http://journal.frontiersin.org/article/10.3389/fgene.2013.00094/full
Also look at this: Blood Cell Origin of Circulating MicroRNAs: A Cautionary Note for Cancer Biomarker Studies
Colin C. Pritchard, Evan Kroh, Brent Wood, Jason D. Arroyo, Katy J. Dougherty, Melanie M. Miyaji, Jonathan F. Tait and Muneesh Tewari
DOI: 10.1158/1940-6207.CAPR-11-0370 Published March 2012
Hope this helps. Please provide more details if your question hasn't been answered yet. Good luck ! Thanks
Hi Jaynish,
Thank you very much for helping me! I have read your study, and I would like to congratulate to you!Its perfect,and your work has been very implement for me!
I know that the best way to determine hemolysis in samples, is using the miR ratio,but this method is a little hard when you have many samples.
miRnas I am studying, aren't described yet,but I will take a look your papers! Really,the problem of bilirrubine and lipemic samples are very important for me, because they cause an important interference in haemolysis at 414nm. I keep working on it.
Thank you very much!!!
María
Thank you Maria for your kind words and good luck with your project :) :)
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